Phenyl propionate of testosterone



2,755,292 PHENYL PROPIONATEDF TESTOSTE O f Colin Leslie Hewett, Glas ow, Scotland, .fliSigDDI vto .0;- ganon Laboratories Limited, London, Englsnd,a'B1-1tish company N .Dmw g- App ca ion. Janua y .3, 119. 3,

Serial.No. 331,.1,16

Claims priority, application Great'Britain January 21, 1952 1 Claim. (.Cl. 2 60-8914) This invention relates to .new derivatives of compounds of the cyclopentanqpolyhydrophenanthrene series and.

compounds having a free :hydroxyl group .capable of -beingaesterified withya fatty acid, are capableofzreacting with .fl-phenyl :propionic acid to physiologically active .the hitherto iknOWn derivatives of the respective'active compounds and .in addition in mosticasesdiaveaihigher degree of activity than the-knownderivatives.

As examples of .these new fiephenyl propionates-smay :be mentioned the mom, di-esters.-of I$7PhHYLPIOPiOHiC acid and oest-radiol, -17.-ethinyl ,oest-radioi, l7methyl .androstenediol-SJY, A 3,21- dihy.droXy .p'regnene tone, .oestrone, .1l-desoxycorticosterone, 17'-hyd-roxy 1 1=desoxy- 'corticosterone (Reichsteins substance ;S) :1=7-hytlroxy corticosterone (Kendallf-s .These esters have not been described {in :literature as .yet. .Theyare capable of. -exerti ng a physiologicaleaction for -a.;considerably long period of time and in many cases i have a higher degree of activity than the known-hitherto used derivatives of the .various compounds. This carries with it the great advantage that less injections are needed, while the high degreeof activity renders-it possible to maintain a therapeutical effect with a much lower dosage.

'The new ,Bfi-phenyl propionates can be prepared e.

by condensati-ng the steroid hormone .withgfi-p nyl propionic acid anhydride, either without. using the.; solven t,

*thereaction time being capable of beingshortenedJ heating or in a solvent, e. g. pyridine, benzene or another suitable solvent or a mixture hereof. 7 ey a .a1 s prep r bymean o g-p teny propionic acid chloride'in'a solvent, suchas pyr'idineor aminture-of benzene and pyridine, which jshould contain As u h are app e th compound F), cortisone, testosterone, 17-ethinyl testosterone, 17 -m ethy1 itestost'er- .one.

nited States Patent 0 0 rderivatives, whichhavea longer duration:of;activity than adequate pyridine to bind the split ofl hydrochloric acid. the sqmman y use me o s f the Prep t o esters of icarboxylic acids may also be applied here, such as theconversion of a diazo ketone with a carboxylic acid.

' The .following examples illustrate several methods for the preparation of these new esters.

iEaa-mnl T s 0sr r n -fi-phe y p opia 'A'solution of 10 gms. of testosterone in cc. of pyridine is cooled in ice and 7.5 gms. of p pheny1- propionyl chloride added slowly with stirring. The solution is then removed from the ice bath, "left at room temperature overnight and then decomposed with ice. The solid ester is recrystallized from methanol; melting point ll61,17 C. (0t D q=+88 (c= 1. in diox ane.

Example II .Asolution of .10 gms. of testosterone in 30 cc. benzene andI'OccU yridine is cooled inice and 7. 5 gms. fl-phenyl- .propion'yl chloride added dropwise with stirring. The solution :is then removed from the ice bath, left overnight-at room-temperature and then washed with water, .di lute sodium carbonate .solution and then dilute hydro- .chloric acid solution. The benzene solution after drying sis .distilled :to dryness and recrystallized from methanol; .meltingspoint v11 6-117 C. I

. Exa pl .11 To a solution of 10 gms. of testosterone ;in .30 cc. pyridine is added 12.5,gms. of B-phenylpropionic anhydride. After standing overnight at room temperature the solution is decomposed by addition of water, and the ;t;estosterone phenyl-propiomte crystallized .from methanol; :meltin lpoin 1165-117 Example lV l0 gms. of testosterone and 12.5 gms. of .p-phenyl propionic anhydride are heated together at 100 C. for fiv'e hours. Themelt is then triturated with water and thesplid washed with dilute sodiumcarbonate. solution, .dtie .ansis ys' all d m hanol. m ep n 1 6- fl he following table ,gives a comparison of 3 the properties of the new testosterone derivatives according to :the inventi n, w t h eady known tQ t .-PrQP 9 a derivatives. Ihetable shows the mean seminal vesicle .we'ights (in rngs) at various intervals after .a single .inter- .mus ulsri ict n 0f 0. t a$t t f$ th u'lt J a ra o mpa t v i qn t be n give'n. The ,different injections listed are prepared as follows: H

Injectioni: flfestosteronepropionate in sesame ,oil 2.5

mgm. in 0.1 cc. oil. 7

injection .IIi Testosterone phenyl propionate in sesame oil32.5-.mgm. in 0.1 cc. oil.

Inject-ion .111": Testosterone propionate in ethyl oleate 2:5mgrn.:inz().1 cc. of the oleate.

:Lnjection .IV: Testosterone phenyl propionate in ethyl toleate:0.-5mgm. in 0.1 cc. of-theoleate.

injection .V: Testosterone phenyl propionate in ethyl oleate 2.5 -'mgm. in 0.1 cc. of the oleate. l

', TAB L'E Jadays 9-.days .14days 21 days 28days days 42days i 1 in Q o It will be noted from the foregoing table that with Injection I maximum activity is obtained in seven days, whereas with Injection II maximum activity is reached in nine days with a much greater degree of activity; moreover even after 42 days the activity is still in general much higher than is reached at any time with Injection I. Comparison of Injections III and IV shows that the activity of the new derivative (Injection IV) is such that the fraction (one-fifth) of the active component present in the Injection IV compared with Injection III gives approximately the same results as with the propionate of Injection III. Comparison of the Injections III and V shows that with the same quantity of active material a still higher peak of activity is reached after fourteen days and that a great measure of activity is preserved even after 42 days.

Example A mixture of 40 gms. of 21-diazopregnenolone and 130 gms. of phenyl propionic acid was heated on a steam bath for 1 hour. The evolution of nitrogen had almost ceased after 25 minutes. The mixture which solidified on cooling, was allowed to stand 16 hours, dissolved in warm methanol, cooled rapidly and poured with stirring into an aqueous solution of sodium carbonate. The precipitate was collected by filtration and washed with warm water until the filtrate reacted alkali free to litmus. The dried crude material (41.5 gm.) was crystallized from methanol to give a faintly yellow amorphous solid, M. P. 108-109.5 C.; (a)n =+31.5: yield 35.2 gm. After recrystallization from methanol the product had M. P. 110.5-112 C.

Example VI.I] desoxycorticosterone 21 B-phenylpropionate Pregnenediolone 21-,9-phenylpropionate (34 gm.) was dissolved in dry toluene (1110 cc.) and cyclohexanone (178 cc.) and the solution distilled until the distillate was clear. Aluminum iso propoxide (8.1 gm.) dissolved in dry toluene (205 cc.) was added and the solution distilled under reflux for 45 minutes. The solution was then cooled rapidly and shaken for 20 minutes with a solution of Rochelle salts (21.9 gm.) in water (44 cc.). The mixture was steam distilled for 80 minutes to remove the toluene and cyclohexanone, cooled and the resulting sticky solid (30 gm.) collected by filtration and washed with a little warm water. The crude solid was dissolved in benzene (100 cc.) and filtered through a column of alumina. The benzene was distilled off and the residue crystallized from methanol. The solid was collected and washed with a little diethyl ether to give a colorless product. crystals (a)r) =+170.5. 139-139.5 C.

Example VII.Oestradiol-3-B-phenylpropionate Oestradiol (13 gm.) with M. P. 178 C. was dis- The product had M. P. of

G solved in warm pure dry acetone (104 cc.) and sodium hydroxide solution (24.7 gm. in 260 cc. distilled water) added to the solution. The solution was then cooled to room temperature. B-Phenyl-propionyl chloride (16 cc.: freshly prepared and distilled) and sodium hydroxide solution (6.5 cc. of 18 gm. dissolved in 25 cc. distilled water) were added simultaneously over 1% hours with vigorous stirring. Stirring was continued for 45 minutes to ensure that all the acid chloride was decomposed. The

Recrystallization from methanol gave colorless ester was collected by filtration and after washing with sutficient 30% acetone to remove the mother liquors, was washed with cold distilled water and finally with warm water until the filtrate reacted as alkali free to red litmus;

The ester at this stage weighed 14.8 gm.-

The mother liquors were then treated as above with fl-phenylpropionyl chloride (8 cc.) and sodium hydroxide solution (3.25 cc.) when a second small crop (2.6 gmf) 4 was obtained. Further treatment gave a third crop (1.2 gm.) The three crops were bulked and crystallized from a. mixture of diethyl ether and n-hexane when the ester was obtained as colorless crystals (16 gm.), M. P. 90.5- 92 C.; (a)r =+62.7. A specimen crystallized from isopropyl ether had M. P. l04.5-l05.5 C.

Example VIII .Oestradiol-3 I 7-di-B-phenylpropionate fl-Phenylpropionyl chloride (30 cc.) was added slowly and with cooling to a solution of a Oestradiol (10 gm.) with M. P. 178 in pyridine cc.). The cherry red solution was allowed to stand 16 hours at room tempera ture when it was poured into a mixture of dilute hydrochloric acid solution and ice. The gummy solid which separated, was dissolved in diethyl ether and washed with sodium carbonate solution, water, dilute hydrochloric acid and finally with water. The ethereal solution, after drying over sodium sulphate was evaporated to dryness and the residue crystallized from acetone to give colorless crystals (9.8 gm.) M. P. -127 C.

Concentration of the filtrate gave a second crop (2.8 gm.). The mother liquors were evaporated to dryness, the residue dissolved in benzene and filtered through a column of alumina. Removal of the benzene by distillation followed by crystallization of the residue from acetone gave colorless crystals (1.4 gm.) M. P. 122-125 C.

The first crop after one further crystallization from acetone had M. P. -130.$ C.; (a)n'=+38.7 in dioxane.

Example IX .-Oestradiol-I 7-B-phenylpr0pi0nate A suspension of Oestradiol-3:l7-di-fl-phenylpropionate (6 gm.) in 0.5% potassium carbonate solution in 95% methanol (1260 cc.) was stirred continuously for 7 hours. The small quantity of undissolved di-ester was removed by filtration and the filtrate neutralized on phenolphthalein with 10% acetic acid solution. The solution was concentrated to approximately 100 cc., diluted with water (400 ml.) and extracted with di-ethyl ether. The ethereal extract was washed with dilute hydrochloric acid solution, water, sodium carbonate solution and finally with water before drying over sodium sulphate. The ethereal solution was concentrated to approximately 25 cc., n-hexane added until slight turbidity resulted the sides of the flask scratched until the ester crystallized when a little more n-hexane was added and the mixture chilled at 0 C. for several hours. The 17-B-phenylpropionate (3.9 gm.) had M. P. 119-121 C. and after one recrystallization from a mixture of diethyl ether and n-hexane it had M. P. 1l9.5-121.5 C.; (a) :+54.5 in dioxane.

Example X .A] 7 rat-methyl 3,9,1 7 fl-dihydroxy androstene- 3-fl-phenylpropionate To a solution of 10 gm. of 17 a-methyl androstenediol in 20 cc. of pyridine are added 45 cc. of absolute benzene. The solution is cooled to 10 C. and to this is added dropwise, while stirring vigorously, a solution of 7 gm. of fl-phenyl propionic acid chloride in 20 cc. of absolute benzene. The reaction mixture is stirred another 15 minutes and cooled down 24 hours at about 0 -5 C. Then it is poured out on a mixture of ice and water. The benzene layer is separated and then washed with 5% sulphuric acid (to acid reaction), with 5% sodium hydroxide solution and with water to neutral reaction. The benzene solution is dried on sodium sulphate, after which it is evaporated to small volume and then filtered through a column of 50 gm. of aluminum oxide. The resulting benzene eluates are evaporated to dryness. After recrystallizing the drying rest from methanol, one obtains 10 gm. of ester with M. P. 1225-1245 C. (ud1)"- -47 (c=1 .0 in dioxane).

Example XI.17-ethtnyl oestradiol 3-p-phenylproplanate 10 of ethinyl 088111111101 dissolved in 40 cc. of pyridine and 10 cc. of benzene. To this solution which has been cooled down to 10 C., is added dropwise, while stirring vigorously, a solution of 8 gm. of fi-phenyl propionic acid chloride in 20 cc. of benzene. After 24 hours standing at -5 C. the reaction mixture is poured out into a mixture of ice and water. The benzene layer, after separating, is washed with sulphuric acid solution, a 5% sodium hydroxide solution and with water. This solution, after drying on sodium sulphate, is evaporated to small volume and filtered through aluminum oxide. The benzene eluates are evaporated to dryness; after recrystallizing from benzene-petroleum ether, the drying rest yields 11 gm. of the ester with M. P. 105- 106 C.; (oc) =3.3 (c=0.9 in dioxane).

Example XII ydr0xy-11-des0xycortic0ster0ne, 21-

fi-phenyl propionate To a solution of 5 gm. of 17-hydroxy-1l-desoxycorticosterone in 25 cc. of pyridine, which is cooled down to C., is added dropwise, while stirring vigorously, a solution of 5 gm. of ,B-phenyl propionic acid chloride in 20 cc. of dried benzene. The reaction mixture is cooled for 24 hours at a temperature of 0-5 C. After pouring out the reaction mixture on a mixture of ice and water, the benzene layer is separated and then washed with 5% sulphuric acid solution, a 5% sodium hydroxide solution and with water. The benzene solution is then dried in sodium sulphate, evaporated to small volume and filtered through 25 gm. of aluminum oxide. The ester is eluded with ether and the resulting solution is evaporated to dryness. After recrystallization from methanol one obtains 5 gm. of the ester with M. P. l70172 C.; (00 +118 (c=0.8 in dioxane).

Example XIII.-Oestr0ne 3-fl-phenylpropionate A solution of 10 gm. of oestrone in 40 cc. of pyridine is cooled down to 10 C. To this is added dropwise, while stirring vigorously, 10 gm. of fl-phenyl propionic acid chloride in 20 cc. of dried benzene. After 24 hours standing at 0-5 C. the reaction mixture is poured out on a mixture of ice and water. The benzene layer, after separating, is washed with a 5% sulphuric acid solution, then with 5% sodium hydroxide solution and with water. The benzene solution is dried on sodium sulphate and filtered through a column of 50 gm. of aluminum oxide. The benzene eluates are evaporated to dryness. The dry- 6 ing rest is recrystallized from acetone, which yields 12.4 gm. of the ester with M. P. 146-148 C.; (a) =+ll1 (c=0.9 in dioxane).

Example XIV.11-dehydro 1 7-hydroxy-corticosterone 21 -fl-phenylpr0pi0nate 3 gm. of cortisone are dissolved in 40 cc. of pyridine. To this, after cooling down to 10 C., is added dropwise, while stirring vigorously, a solution of 3 gm. of ,8- phenyl propionic acid chloride in 10 cc. of benzene. The reaction mixture is left to stand for 24 hours at 05 C. and then poured out into a mixture of ice and water. The benzene layer is separated and then washed with a 5% sulphuric acid solution, then with 5% sodium hydroxide solution and with Water. The benzene solution, after drying on sodium sulphate, is evaporated to small volume and filtered through a column of 15 gm. of aluminum oxide. The ester is eluted with a mixture of benzene and ether (:50), then the eluates are evaporated to dryness. Finally the drying rest is recrystallized from a mixture of acetone and ether. Obtained are 3 gm. of the fl-phenyl propionate with M. P. -177 C. (00 +183 (c=0.5 in dioxane).

Example X V.A 11 .17.21-zrihydr0xy-3,20-diketo pregnene 21 -fi-phenyl propionate To a solution of 200 mg. of compound F (Kendall) in 2 ml. of pyridine are added 0.2 ml. of fi-phenyl propionic acid anhydride. The reaction mixture is left to stand overnight at room temperature. The ester is crystallized from aqueous acetone. The crystals have a M. P. of 138139 C. (a) =+136 (c=0.7 in dioxane).

What I claim is:

The biologically active B-phenyl propionate of testosterone with M. P. 116-1 17 C.

References Cited in the file of this patent UNITED STATES PATENTS 2,109,400 Miescher Feb. 22, 1938 2,156,599 Miescher May 2, 1939 FOREIGN PATENTS 208,892 Switzerland June 1, 1940 

1. THE BIOLOGICALLY ACTIVE B-PHENYL PROPIONATE OF TESTOSTERONE WITH M. P. 116-117* C. 